Methods for diagnostic screening

Abstract

Methods are presented for mass screening of patient populations for indicia of disease, infection, or predisposition to disease.

What is claimed is:  
     
         1 . A method for diagnosing the disease state of a patient, the method comprising the steps of: 
 (a) combining tissue or body fluid samples obtained from a plurality of patients, thereby to form a combined sample;    (b) analyzing said combined sample for the presence of a disease marker; and    (c) diagnosing the disease status of each member of said plurality by conducting at least one step selected from the group consisting of: 
 (1) Identifying each member of said plurality as being healthy if no disease marker is detected in said analyzing step; and  
 (2) Serially analyzing patient samples if a disease marker is detected in said analyzing step; 
 thereby to diagnose the disease state of each member of said plurality.  
 
   
     
     
         2 . The method of    claim 1   , wherein said disease marker is a nucleic acid.  
     
     
         3 . The method of    claim 2   , wherein said nucleic acid is a mutation.  
     
     
         4 . The method of    claim 3   , wherein said analyzing step comprises 
 (a) exposing said combined sample to a first nucleic acid probe capable of specific hybridization with a nucleic acid known or suspected to be mutated in diseased cells,    (b) exposing said combined sample to a nucleic acid probe capable of specific hybridization with a nucleic acid known not to be mutated in disease cells;    (c) enumerating a number of first and second probes that hybridize in said combined sample; and    (d) determining whether a statistically-significant difference exists between the number of first and second probes.    
     
     
         5 . The method of    claim 2   , wherein said analyzing step comprises the steps of 
 (a) annealing an oligonucleotide primer to a nucleic acid sample under conditions that promote exact complementary hybridization between said primer and a portion of a nucleic acid in said combined sample;    (b) extending said primer by a single base; and    (c) identifying said single base.    
     
     
         6 . The method of    claim 5   , further comprising the step of determining whether said single base is a known polymorphic variant indicative of disease.  
     
     
         7 . The method of    claim 2   , wherein said analyzing step comprises 
 (a) exposing said combined sample to a nucleic acid primer under conditions that promote hybridization of said probe to a nucleic acid region immediately downstream of a single nucleotide polymorphic locus;    (b) exposing said sample to at least four different chain-terminating nucleic acids under conditions that promote extension of said primer;    (c) isolating extended primer in either from primer that has not been extended;    (d) determining a number of each unique chain terminating nucleic acid attached to an extended primer; and    (e) determining if a statistically-significant difference occurs between said numbers.    
     
     
         8 . The method of    claim 1   , wherein said tissue or body fluid sample is selected from the group consisting of sputum, stool, blood, cerebrospinal fluid; biopsy tissue, urine, semen, lymph, and pap smear.  
     
     
         9 . The method of    claim 1   , wherein said disease is selected from the group consisting of cancer, diabetes, amyotropic lateral sclerosis, AIDS, Alzheimer's disease, and parasitic diseases.  
     
     
         10 . The method of    claim 1   , wherein said plurality comprises from 2 to about 25 patients.  
     
     
         11 . The method of    claim 1   , wherein said plurality comprises 100 patients.  
     
     
         12 . The method of    claim 1   , wherein said plurality comprises 1000 patients.  
     
     
         13 . The method of    claim 3   , wherein said mutation is selected from the group consisting of a point mutation, loss of heterozygosity, a rearrangement, a deletion, and inversion, and a translocation.  
     
     
         14 . The method of    claim 1   , wherein said DNA is isolated from said combined sample prior to said analyzing step.